姜黄素是从姜黄植物中提取出的一种抗增殖植物化学物质，一直以来在细胞单层和动物模型的临床前药物筛选中得到研究。然而，这些培养系统存在一些限制，通过进行三维细胞培养筛选可以克服这些限制。本研究的目的是通过对宫颈腺癌HeLa细胞球体进行RT-PCR基因表达分析（包括参与细胞死亡的CASP3、CASP8、CASP9、PARP1、BBC3、BIRC5、BCL2、TNF基因，参与自噬的BECN1、SQSTM1基因，参与细胞周期调节的TP53、C-MYC、NF-kB、CDKN1A、m-TOR、TRAF-2基因，参与DNA损伤修复的H2AFX、GADD45A、GADD45G基因，参与氧化应激的GPX1基因，参与内质网应激的EIF2AK3、ERN1基因，以及参与侵袭的MMP1、MMP9基因）来研究姜黄素对细胞毒性、基因毒性和球体生长的影响。姜黄素对细胞具有浓度依赖性的细胞毒性作用。姜黄素处理的细胞球体在克隆形成实验中表现出较低的增殖恢复和细胞增殖减弱。另外，未观察到明显的基因毒性。姜黄素处理的细胞球体显示BECN1 (2.9倍)、CASP9 (2.1倍) 和PARP1 (2.1倍) mRNA表达降低。PARP1的抑制暗示了增殖维持的关键途径的破坏。CASP9 mRNA表达的下调和CASP3 / 8 mRNA表达的不变表明非半胱天冬酶依赖性细胞死亡，而BECN1 mRNA的下调则表示自噬的破坏。因此，姜黄素具有作为抗增殖活性药物开发的潜力，可以考虑在癌症治疗中使用。

Curcumin is an antiproliferative phytochemical extracted from Curcuma longa L and which has been studied in preclinical drug screening using cell monolayers and animal models. However, several limitations of these culture systems may be overcome by performing screening with three-dimensional (3-D) cell culture. The aim of this study was to investigate the effects of curcumin on cytotoxicity and genotoxicity as well as spheroid growth using cervical adenocarcinoma HeLa cell spheroids by performing RT-PCR mRNA expression of genes involved in cell death (CASP3, CASP8, CASP9, PARP1, BBC3, BIRC5, BCL2, TNF), autophagy (BECN1, SQSTM1), cell cycle regulation (TP53, C-MYC, NF-kB, CDKN1A, m-TOR, TRAF-2), DNA damage repair (H2AFX, GADD45A, GADD45G), oxidative stress (GPX1), reticulum stress (EIF2AK3, ERN1), and invasion (MMP1, MMP9) was investigated. Curcumin was cytotoxic in a concentration-dependent manner. Curcumin-treated spheroids exhibited lower proliferative recovery and cell proliferation attenuation, as observed in the clonogenic assay. Further, no marked genotoxicity was detected. Curcumin-treated spheroids displayed reduced expression of BECN1 (2.9×), CASP9 (2.1×), and PARP1 (2.1×) mRNA. PARP1 inhibition suggested disruption of essential pathways of proliferation maintenance. Downregulated expression of CASP9 mRNA and unchanged expression of CASP3/8 mRNA suggested caspase-independent cell death, whereas downregulated expression of BECN1 mRNA indicated autophagic disruption. Therefore, curcumin exhibits the potential for drug development with antiproliferative activity to be considered for use in cancers.